Provided are a thin-film transistor substrate that has enhanced electrical characteristics, such as off-current characteristics of a thin-film transistor, without increasing the number of mask processes, a display apparatus, and a method of manufacturing the thin-film transistor substrate. The thin-film transistor substrate includes: a semiconductor layer including a first conductive region, a second conductive region, and a first semiconductor region; a lower electrode disposed on the semiconductor layer and at least partially overlapping the first semiconductor region; and an upper electrode disposed on the lower electrode and at least partially overlapping the first semiconductor region, a first boundary between the first semiconductor region and the first conductive region coincides with an edge of the upper electrode, and a second boundary between the first semiconductor region and the second conductive region coincides with an edge of the lower electrode or an edge of the upper electrode.